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Effects of a Modified Dye-Labeled Nucleotide Spacer Arm on Incorporation by Thermophilic DNA Polymerases

Lacenere, Christopher J. and Garg, Neil K. and Stoltz, Brian M. and Quake, Stephen R. (2006) Effects of a Modified Dye-Labeled Nucleotide Spacer Arm on Incorporation by Thermophilic DNA Polymerases. Nucleosides, Nucleotides and Nucleic Acids, 25 (1). pp. 9-15. ISSN 1525-7770. doi:10.1080/15257770500377714.

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The ability of eight commercially available thermophilic DNA polymerases to sequentially incorporate fluorescently labeled nucleotides sequentially was analyzed by a gel based primer extension assay. Cy5-dUTP or a variant nucleotide in which the linker had been lengthened by 14 atoms between the dye and the nucleobase were compared. We found that the Cy5-dUTP with a longer linker resulted in longer primer extension lengths. Furthermore, some of the assayed polymerases are capable of extending the primer to the full or near full length of 30 nucleotides using dye-labeled nucleotides exclusively.

Item Type:Article
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URLURL TypeDescription
Stoltz, Brian M.0000-0001-9837-1528
Quake, Stephen R.0000-0002-1613-0809
Additional Information:© 2006 Taylor & Francis Group, LLC. Received 19 February 2005; accepted 26 May 2005. This work was supported in part by DARPA, the National Institutes of Health, and the Rosen fellowship. The authors would like to thank Peter Dervan and his group for the use of their imager. C. L. thanks John DeModena and Saurabh Vyawahare for their useful advice and comments.
Funding AgencyGrant Number
Defense Advanced Research Projects Agency (DARPA)UNSPECIFIED
Rosen FellowshipUNSPECIFIED
Subject Keywords:DNA polymerase, Fluorescent nucleotide, Sequencing by synthesis
Issue or Number:1
Record Number:CaltechAUTHORS:20170223-112338543
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:74497
Deposited By: Ruth Sustaita
Deposited On:23 Feb 2017 20:53
Last Modified:11 Nov 2021 05:27

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